The Coenzyme Q10 (CoQ10) BioAssay ELISA Kit is a quantitative competitive assay for the detection of CoQ10 in human serum,plasma,tissue homogenates and other biological fluids.Detection Range:0.781-50ng/mlSensitivity:<0.469ng/mlPrecision:Intra-assay CV: <8%Inter-assay CV: <10%Assay Principle:This ELISA kit employs the quantitative competitive enzyme-linked immunoassay technique,utilizing a microtiter plate provided in this kit has been pre-coated with CoQ10. Standards and samples are added to the appropriate microtiter plate wells followed by a biotin-conjugated antibody specific to CoQ10. Streptavidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added,only those wells that contain CoQ10,Biotin-conjugated antibody and enzyme-conjugated Streptavidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulfuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of CoQ10 in the sample is then determined by comparing the O.D. of the sample to the standard curve.Kit Components:*369832A: Microtiter Strips,1x96 wells (8x12 wells).*369832B: Standard,2x1 vials369832C: Sample/Standard Dilution Buffer,1x20ml369832D: Antibody (Biotin) (Concentrated),1x60ul369832E: Antibody Dilution Buffer,1x10ml369832F: Streptavidin (HRP) (SABC),1x120ul369832G: SABC Dilution Buffer,1x10ml369832H: TMB Substrate,1x10ml369832J: Stop Solution,1x10ml369832K: Wash Buffer,25X,1x30mlStorage and Stability:Store unopened *369832A at 4ºC; store at -20ºC once opened. Store unopened *369832B at 4°C; once reconstituted,store at 4°C for up to 12 hours or at -20°C for up to 48 hours. Store other components at 4°C. Kit is stable for 6 months after receipt. For maximum recovery of product,centrifuge the original vials after thawing and prior to removing the cap.Assay Summary:1. Wash plate 2 times before adding standards and samples to wells.2. Add 50ul standard or sample to each well.3. Immediately add 50ul Antibody-Biotin working solution to each well and incubate for 45 minutes at 37°C4. Aspirate and wash 3 times.5. Add 100ul SABC working solution to each well. Incubate for 30 minutes at 37°C6. Aspirate and wash 5 times.7. Add 90ul TMB substrate. Incubate 15-20 minutes at 37°C8. Add 50ul Stop Solution. Read at 450nm immediately.9. Calculate results.